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. 2012 Oct 9;287(50):41808–41819. doi: 10.1074/jbc.M112.376558

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 7. — Effect of CDKAL1 overexpression on insulin granule protein content. A, INS-1 cells were transfected by electroporation with different amounts of CDKAL1-HA3. 72 h after transfection, cell extracts were analyzed by immunoblot for the indicated granule proteins. As a short exposure with anti-CDKAL1 Ab1 is shown, endogenous CDKAL1 is not visible. γ-Tubulin was used as loading control. B, rescue of CDKAL1 silencing by overexpression of CDKAL1-HA3. INS-1 cells were transfected with CDKAL1-HA3 or the empty plasmid and 1 day later with CDKAL1-siRNA or control-siRNA. 48 h after transfection of siRNA oligo, cell extracts were analyzed by immunoblot for the indicated granule proteins. Note that CDKAL1-HA3 is not targeted by the specific siRNA oligos. Overexpression of CDKAL1-HA3 in the context of CDKAL1 silencing (lane 4) counteracts the decrease of the three proteins examined. γ-Tubulin was used as loading control.